STED-fluorescent light microscopy with two-photon excitation

US7863585B2 · kind B2 · utility

Assignee

• Max-Planck-Gesellschaft zur Föderung der Wissenschaften e.V. · US

Inventors

• Stefan W. Hell · Oebisfelde, DE
• Katrin Willig · Oettingen, DE

Key dates

• Filing date: Feb 16, 2010
• Grant date: Jan 4, 2011
• Expiry date: Feb 16, 2030

Classification

• Technology area (CPC G): Physics
• CPC primary: G02B21/0076
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

A method of high spatial resolution imaging a structure in a sample comprises: marking the structure with molecules of a fluorescent dye; selecting a first wavelength for excitation light which excites the molecules of the fluorescent dye via a multi photon process for spontaneous emission of fluorescent light; focusing pulses of the excitation light into the sample to excite those molecules of the fluorescent dye present in a focal area of the focused excitation light; selecting a second wavelength shorter than the first wavelength for de-excitation light which de-excites excited molecules of the fluorescent dye prior to their spontaneous emission; during a plurality of the pulses of the excitation light, continuously directing the de-excitation light onto the sample to de-excite excited molecules of the fluorescent dye, which are located outside an measurement area which is a fraction of the focal area; and recording the fluorescent light spontaneously emitted by the molecules of the fluorescent dye in the sample.